LOCUS       sequence-309649-        6214 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses membrane localized tagBFP with wrmScarlet11 C-terminal 
            fusion in S. cerevisiae.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6214)
  AUTHORS   Goudeau J, Sharp CS, Paw J, Savy L, Leonetti MD, York AG, Updike DL,
            Kenyon C, Ingaramo M
  TITLE     Split-wrmScarlet and split-sfGFP: tools for faster, easier 
            fluorescent labeling of endogenous proteins in Caenorhabditis 
            elegans.
  JOURNAL   Genetics. 2021 Apr 15;217(4). pii: 6126424. doi: 
            10.1093/genetics/iyab014.
  PUBMED    33693628
REFERENCE   2  (bases 1 to 6214)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Genetics.";
            date: "2021-04-15"; volume: "217(4). pii: 6126424. doi"; pages: " 
            10.1093/genetics/iyab014"
COMMENT     Sequence Label: p416-TEF-membrane localization
            signal-tagBFP-w...
FEATURES             Location/Qualifiers
     source          1..6214
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             91..789
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK
                     LVGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC
                     DLPSKLGHKLN"
     terminator      883..1080
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(1094..1113)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1095..1113)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1123..1140)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1123..1139)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1132..1154)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      1280..1735
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1367..1386)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1577..1598
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(1866..2669)
                     /codon_start=1
                     /gene="S. cerevisiae URA3"
                     /product="orotidine-5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=URA3
                     /note="yeast auxotrophic marker, counterselectable with 
                     5-fluoroorotic acid (5-FOA)"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     promoter        complement(2670..2885)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(2909..2928)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3028..3050
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3088..3106)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    3147..3650
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        3687..3791
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3792..3860,3861..4652)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(4010..4029)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4823..5411
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5312..5331
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5565..5582
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5699..5720
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5735..5740,5741..5758,5759..5765)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5773..5789
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     5778..5800
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5797..5813
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5797..5813
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     5832..5852
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        5834..5852
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        5871..6214
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
ORIGIN
        1 atgggtttta tatcatctat cctatgttgc agctctgaaa ctacgcagag taactccaac
       61 tcagcgtata ggcaacagca atccactagt atgagcgagc tgattaagga gaacatgcac
      121 atgaagctgt acatggaggg caccgtggac aaccatcact tcaagtgcac atccgagggc
      181 gaaggcaagc cctacgaggg cacccagacc atgagaatca aggtggtcga gggcggccct
      241 ctccccttcg ccttcgacat cctggctact agcttcctct acggcagcaa gaccttcatc
      301 aaccacaccc agggcatccc cgacttcttc aagcagtcct tccctgaggg cttcacatgg
      361 gagagagtca ccacatacga agacgggggc gtgctgaccg ctacccagga caccagcctc
      421 caggacggct gcctcatcta caacgtcaag atcagagggg tgaacttcac atccaacggc
      481 cctgtgatgc agaagaaaac actcggctgg gaggccttca ccgagacgct gtaccccgct
      541 gacggcggcc tggaaggcag aaacgacatg gccctgaagc tcgtgggcgg gagccatctg
      601 atcgcaaaca tcaagaccac atatagatcc aagaaacccg ctaagaacct caagatgcct
      661 ggcgtctact atgtggacta cagactggaa agaatcaagg aggccaacaa cgagacctac
      721 gtcgagcagc acgaggtggc agtggccaga tactgcgacc tccctagcaa actggggcac
      781 aagcttaatg gcggggggtc ttataccgtt gtcgagcaat atgaaaagtc agtggcgagg
      841 cactgtacag ggggataaga cgagctatat aagtaactcg agactgacaa taaaaagatt
      901 cttgttttca agaacttgtc atttgtatag tttttttata ttgtagttgt tctattttaa
      961 tcaaatgtta gcgtgattta tatttttttt cgcctcgaca tcatctgccc agatgcgaag
     1021 ttaagtgcgc agaaagtaat atcatgcgtc aatcgtatgt gaatgctggt cgctatactg
     1081 ggtacccaat tcgccctata gtgagtcgta ttacgcgcgc tcactggccg tcgttttaca
     1141 acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat cgccttgcag cacatccccc
     1201 tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg
     1261 cagcctgaat ggcgaatgga cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     1321 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
     1381 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     1441 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
     1501 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     1561 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
     1621 gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag
     1681 ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgcttac aatttcctga
     1741 tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcataggg taataactga
     1801 tataattaaa ttgaagctct aatttgtgag tttagtatac atgcatttac ttataataca
     1861 gttttttagt tttgctggcc gcatcttctc aaatatgctt cccagcctgc ttttctgtaa
     1921 cgttcaccct ctaccttagc atcccttccc tttgcaaata gtcctcttcc aacaataata
     1981 atgtcagatc ctgtagagac cacatcatcc acggttctat actgttgacc caatgcgtct
     2041 cccttgtcat ctaaacccac accgggtgtc ataatcaacc aatcgtaacc ttcatctctt
     2101 ccacccatgt ctctttgagc aataaagccg ataacaaaat ctttgtcgct cttcgcaatg
     2161 tcaacagtac ccttagtata ttctccagta gatagggagc ccttgcatga caattctgct
     2221 aacatcaaaa ggcctctagg ttcctttgtt acttcttctg ccgcctgctt caaaccgcta
     2281 acaatacctg ggcccaccac accgtgtgca ttcgtaatgt ctgcccattc tgctattctg
     2341 tatacacccg cagagtactg caatttgact gtattaccaa tgtcagcaaa ttttctgtct
     2401 tcgaagagta aaaaattgta cttggcggat aatgccttta gcggcttaac tgtgccctcc
     2461 atggaaaaat cagtcaagat atccacatgt gtttttagta aacaaatttt gggacctaat
     2521 gcttcaacta actccagtaa ttccttggtg gtacgaacat ccaatgaagc acacaagttt
     2581 gtttgctttt cgtgcatgat attaaatagc ttggcagcaa caggactagg atgagtagca
     2641 gcacgttcct tatatgtagc tttcgacatg atttatcttc gtttcctgca ggtttttgtt
     2701 ctgtgcagtt gggttaagaa tactgggcaa tttcatgttt cttcaacact acatatgcgt
     2761 atatatacca atctaagtct gtgctccttc cttcgttctt ccttctgttc ggagattacc
     2821 gaatcaaaaa aatttcaagg aaaccgaaat caaaaaaaag aataaaaaaa aaatgatgaa
     2881 ttgaaaaggt ggtatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc
     2941 agccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat
     3001 ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt
     3061 catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg cctattttta taggttaatg
     3121 tcatgataat aatggtttct tagacggatc gcttgcctgt aacttacacg cgcctcgtat
     3181 cttttaatga tggaataatt tgggaattta ctctgtgttt atttattttt atgttttgta
     3241 tttggatttt agaaagtaaa taaagaaggt agaagagtta cggaatgaag aaaaaaaaat
     3301 aaacaaaggt ttaaaaaatt tcaacaaaaa gcgtacttta catatatatt tattagacaa
     3361 gaaaagcaga ttaaatagat atacattcga ttaacgataa gtaaaatgta aaatcacagg
     3421 attttcgtgt gtggtcttct acacagacaa gatgaaacaa ttcggcatta atacctgaga
     3481 gcaggaagag caagataaaa ggtagtattt gttggcgatc cccctagagt cttttacatc
     3541 ttcggaaaac aaaaactatt ttttctttaa tttctttttt tactttctat ttttaattta
     3601 tatatttata ttaaaaaatt taaattataa ttatttttat agcacgtgat gaaaaggacc
     3661 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     3721 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     3781 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     3841 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     3901 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     3961 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
     4021 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     4081 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
     4141 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     4201 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     4261 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     4321 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
     4381 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
     4441 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     4501 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     4561 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     4621 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     4681 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     4741 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     4801 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     4861 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     4921 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     4981 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     5041 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     5101 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     5161 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     5221 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     5281 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     5341 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     5401 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     5461 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
     5521 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
     5581 aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt
     5641 aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta
     5701 atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt ccggctcgta
     5761 tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat gaccatgatt
     5821 acgccaagcg cgcaattaac cctcactaaa gggaacaaaa gctggagctc gacatggagg
     5881 cccagaatac cctccttgac agtcttgacg tgcgcagctc aggggcatga tgtgactgtc
     5941 gcccgtacat ttagcccata catccccatg tataatcatt tgcatccata cattttgatg
     6001 gccgcacggc gcgaagcaaa aattacggct cctcgctgca gacctgcgag cagggaaacg
     6061 ctcccctcac agacgcgttg aattgtcccc acgccgcgcc cctgtagaga aatataaaag
     6121 gttaggattt gccactgagg ttcttctttc atatacttcc ttttaaaatc ttgctaggat
     6181 acagttctca catcacatcc gaacataaac aacc
//
